PMID: 37902Jul 10, 1979

Turnover and vectorial properties of cytochrome c oxidase in reconstituted vesicles

Biochimica Et Biophysica Acta
J M Wrigglesworth, P Nicholls

Abstract

1. Proteoliposomes containing cytochrome c oxidase and phospholipid have been made by sonication and by the cholate dialysis procedure. In both methods of preparation, only about 50% of the enzyme molecules are oriented in the membrane with their cytochrome c reaction sites exposed to the outside of the vesicle. 2. The activity of cytochrome c oxidase in the reconstituted vesicles in not increased by incubation in 1% Tween 80. Experiments on reconstituted vesicles containing internal (entrapped) cytochrome c indicate that turnover of enzyme oxidising entrapped cytochrome c in the presence of N,N,N',N'-tetramethyl-p-phenylenediamine or 2,3,5,6-tetramethyl-p-phenylendediamine is at a very much lower rate than enzyme oxidising external ferrocytochrome c. 3. Oxidation of ascorbate by externally added cytochrome c results in an electrogenic production of OH- inside the vesicles, which can be monitored using entrapped phenol red. Polylysine inhibits, but does not abolish, the internal alkalinity change in reconstituted vesicles oxidising internal (entrapped) cytochrome c using externably added ascorbate plus N,N,N',N'-tetramethyl-p-phenylenediamine. When 2,3,5,6-tetramethyl-p-phenylenediamine is used as the permeable redox mediator, ...Continue Reading

References

Feb 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·J R FetterS Ferguson-Miller
May 21, 1980·Journal of Theoretical Biology·Y D Halsey
Oct 15, 1980·Archives of Biochemistry and Biophysics·P NichollsJ M Wrigglesworth
Sep 14, 1981·Biochimica Et Biophysica Acta·L J ProchaskaG Buse
Feb 1, 1982·European Journal of Biochemistry·R P CaseyA Azzi

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Cytochrome c Group
Cholic Acids
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Niosomes
Oxygen Consumption

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